Development and validation of UPLC-PDA method for concurrent analysis of bergenin and menisdaurin in aerial parts of Flueggea virosa (Roxb. ex Willd.)

Bergenin and menisdaurin are biologically active components which are found in plant Flueggea virosa (Phyllanthaceae). Bergenin has pharmacological actions such as chemopreventive and antihepatotoxic while menisdaurin has an anti-viral activity which needs its evaluation by an analytical method (UPLC-PDA method) that can be applied to the quality control of pharmaceutical preparations. The developed UPLC-PDA method was applied for identification and quantification of standards bergenin and menisdaurin in the methanol extract of F. virosa (FVME). The analysis was carried out using Eclipse C18 (4.6?×?100?mm, 3.5?µm) UPLC column. The optimized chromatographic condition was achieved at 0.16?mL/min flow rate using gradient system with acetonitrile and water as mobile phase. Both biomarkers were measured at λ_max 235?nm in PDA detector at ambient temperature. The developed method furnished sharp and intense peaks of menisdaurin and bergenin at Rt?=?2.723 and 3.068?min, respectively along with r2?>?0.99 for both. The recoveries of bergenin and menisdaurin were found in the range of 99.37–101.49% and 98.20–100.08%, respectively. With other validation data, including precision, specificity, accuracy, and robustness, this method demonstrated excellent reliability and sensitivity. The separation parameters i.e. retention, separation, and resolution factors for resolved standards (bergenin and menisdaurin) were >1, which showed good separation. The quantity of bergenin and menisdaurin in the FVME sample was found as 15.16 and 3.28% w/w, respectively. The developed UPLC-PDA method could be conveniently adopted for the routine quality control analysis.     

治咳川贝枇杷滴丸的质量控制及其挥发性成分的测定

采用超高效液相色谱-光电二极管阵列法检测了治咳川贝枇杷滴丸及其相似处方制剂产品的指纹图谱,再用偏最小二乘法进行聚类分析评价研究.结果显示治咳川贝枇杷滴丸过期样品与市售产品间差异最大.继而采用全二维气相色谱-飞行时间质谱(GC×GC/TOF-MS)对治咳川贝枇杷滴丸中的挥发性成分进行定性定量分析.共确定了其中的74个成分,所鉴定的组分占挥发油色谱总馏出峰面积的90.19％.     

UPLC-PDA法同时测定复方五凤草液中7种成分

目的建立UPLC-PDA法同时测定复方五凤草液(五凤草、白芨、猫爪草)中没食子酸、原儿茶酸、对羟基苯甲酸、咖啡酸、金丝桃苷、槲皮素、柚皮素的含有量。方法该药物80%甲醇提取液的分析采用Waters BEH C18色谱柱(2.1 mm×100 mm, 1.7μm);流动相乙腈-0.1%甲酸,梯度洗脱;体积流量0.3 mL/min;柱温40℃;检测波长266 nm。结果 7种成分在各自范围内线性关系良好(r≥0.999 5),平均加样回收率96.96%~102.26%,RSD 0.65%~2.10%。结论该方法稳定准确,重复性好,可用于复方五凤草液的质量控制。     

UPLC-PDA同时测定柴胡中8种多炔类化合物的含量

建立柴胡中多炔类化合物定量测定方法。采用Waters BEH C_(18)柱(2.1 mm×100 mm,1.7μm),甲醇-水溶液为流动相进行梯度洗脱,体积流量0.3 m L·min~(-1),检测波长315 nm,柱温40℃。采用传统提取分离方法,制备得8种多炔类化合物,其中化合物7和8为新化合物。在线性范围内8种多炔类化合物线性良好(r0.999 0);日内精密度和日间精密度均小于1.9%;平均回收率范围在93.21%~108.4%。同时,对收集到的17个柴胡药材样品进行定量测定,发现不同种属、不同产地柴胡中所含多炔类成分种类及含量差异较大。建立的含量测定方法具有良好的精密度、准确度、重复性,可用于柴胡中多炔类化合物的定量测定及质量控制,为选择多炔类成分较多的柴胡药材提供一种简便方法。     

Simultaneous Determination of Eight Constituents in Fruits of Rubus chingii by UPLC

Objective To develop a simple,efficient,and reliable method for routine quantitative analysis of main constituents presented in the fruits of Rubus chingii,which is widely used in Chinese materia medica(CMM),known as Fupenzi(FPZ) in Chinese.Methods An ultra performance liquid chromatography-photo diode array(UPLC-PDA) system was employed for simultaneous quantification of eight compounds,i.e.adenosine,gallic acid,brevifolin carboxylic acid,ethyl gallate,ellagic acid,kaempferol-3-O-rutinoside,kaempferol-3-O-β-D-glucopyranoside,and tiliroside.The chromatographic analysis was performed on a C_(18) column using a gradient elution of acetonitrile-0.1%formic acid aqueous solution within a runtime of 25 min.Results All calibration curves were linear(R~2 0.9997) over the tested ranges.The intra- and inter-day precisions as determined from sample solutions were both less than 2.45%and 2.78%,respectively.The average recoveries for the eight constituents ranged from 94.77%to 101.35%with RSD ≤ 4.41%.The newly-developed method was applied to the quality assessment of various R.chingii samples,including both ripe and unripe fruits of R.chingii from different habitats.Conclusion The relative levels of the investigated compounds vary remarkably in the fruits of R.chingii collected from different habitats.As only two of the eight compounds,adenosine and ellagic acid,are determined in the ripe fruits of R.chingii,the results may explain the reason why only the unripe fruits can be used in CMM.     

Phenological and geographical influence in the concentration of selected bioactive 4-phenylcoumarins and chlorogenic acid in Hintonia latiflora leaves

Ethnopharmacological relevance

Hintonia latiflora is a Mexican medicinal plant with well-documented ethnomedical record comprising more than 400 years; in modern Mexico is used for treating several maladies such as diabetes and gastric ulcers. Although the pharmacological actions of the stem-bark and leaves have been demonstrated, the phenological and geographical effect on the concentration of active principles remains unexplored.

Aim of the study

The main goals of this study were to analyze the amount of selected 4-phenylcoumarins and chlorogenic acid in the leaves in order to assess the best harvesting period, and consequently their pharmacological efficacy. In addition, the preclinical antidiabetic efficacy of the infusion of the leaves was corroborated using standard pharmacological tests.

Materials and methods

The aqueous extracts from the leaves of Hintonia latiflora were prepared by infusion. For phenological and geographical comparison, leaves of Hintonia latiflora were collected in two different regions in Chihuahua and Michoacán. The material was analyzed by UPLC applying an analytical method that developed and validated for this purpose following the ICH guidelines. Investigation of the antidiabetic action was accomplished using an acute hypoglycemic test and oral glucose and sucrose tolerance tests.

Results

The validated analytical method was successfully applied for quantifying chlorogenic acid (1) and 4-phenylcoumarins (2–5) in the leaves of 12 different batches (1–12) during one-year period, and seven different batches for each geographical region; the concentration of the metabolites at the phenological cycle was significantly different, their concentration increased during the pre-senescence phase whereas in the leaf renovation stage the highest concentration of 2–5 was reached. The overall analysis of the active compounds concentration between the two populations investigated seems to be less important than the phenological variations. The aqueous extract of the leaves of Hintonia latiflora exerted its antidiabetic effect by different mechanisms showing comparable effect to the organic extract.

Conclusions

The findings of the present investigation reveal that the best harvest season for the leaves of Hintonia latiflora is between the leaves renovation and senescence stages avoiding the flowering period. In addition, no significant differences were found among the two different geographical populations analyzed. The infusions of the leaves, rich in 4-phenylcoumarins and chlorogenic acid, showed comparable antidiabetic action than the organic extract.     

Development of Ultra-performance Liquid Chromatographic Fingerprint for Quality Control of Rheum palmatum L.

A novel, rapid and valid fingerprint method was developed using ultra-performance liquid chromatography-photodiode array detection （UPLC-PDA） for the quality control of Rheum palmatum L., a traditional Chinese herb with antipyretic, purging and antibacterial actions. For the first time, the feasibility and advantages of employing chromatographic fingerprint were investigated and demonstrated for the quality control of R. palmatum by systematically comparing chromatograms of extracts with the professional analytical software recommended by China＇s State Food and Drug Administration （SFDA）. The results revealed that the chromatographic fingerprint combining similarity evaluation could efficiently identify and distinguish raw herbs of R. palmatum from different species. The effects of different commercial grades on R. palmatum chromatographic fingerprints were also examined.     

In Vitro Stability Evaluation of Different Pharmaceutical Products Containing Meropenem

Background:

Meropenem is a beta-lactam antibiotic for treating multidrug-resistant gram-negative bacilli infections. The expiry of the drug’s patent (Merrem) allowed the production of generics to be commercialized by a few companies, including Hospira and Hikma. The stability of these medicines after reconstitution as reported on a data sheet report is 6 hours for Merrem and 1 hour for generics.

Objectives:

The aim of this work was to evaluate the stability profile of 3 products in 0.9% sodium chloride until 6 hours.

Methods:

Six polyolefin bags (2 for each drug, stored in the light and in the dark) were prepared for every test run (n =10) at concentrations of 4 and 10 mg/mL. All solutions were stored at controlled room temperature (25°C ± 3°C) and sampled immediately after preparation and at every hour until 6 hours had passed. The concentrations, pH changes, and the visual clarity were used as stability and compatibility indicators.

Results:

All 3 drugs retained over 95% of the initial concentration at 3 to 4 hours. At the sixth hour, all the concentrations decayed 8% to 10%. No statistical differences were observed in the percentage deviation values of the stability profile between generics and the branded drug.

Conclusion:

The stability profile of the products in polyolefin bags, at 4 and 10 mg/mL, was superimposable during the period of analysis and seems to show small values of deviation (1%-2%). These data do not affect the pharmacokinetics because these variations could be attributed to the intra- and interindividual variability between patients. The products showed the same stability, and consequently they could be used interchangeably in hospital pharmacy.     

UPLC-PDA同时测定藏药翼首草环烯醚萜苷和酚酸类10个化学成分的含量

建立超高效液相色谱法(UPLC-PDA)同时测定藏药翼首草环烯醚萜和酚酸类10个化学成分的含量。采用Acquity UPLC~R BEH C_(18)(2.1 mm×100 mm,1.7μm)色谱柱,流动相0.2%磷酸水(A)-乙腈(B),梯度洗脱,检测波长237,325 nm,流速0.4 mL·min~(-1),柱温30℃,进样量1μL。研究结果表明UPLC-PDA法定量分析翼首草10个化学成分在考察的范围内线性关系良好(r≥0.999 7);加样回收率为96.30%~103.0%(n=9),RSD为0.72%~2.9%(n=9)。所建立UPLC方法简便、准确、重复性好,可同时测定藏药翼首草中10个化学成分的含量。     

基于UPLC-PDA法逍遥散乙醇提取物的石油醚萃取物血清药物化学初探

目的 明确大鼠ig给予逍遥散乙醇提取物的石油醚萃取物（XYP-A）后血清中移行成分的归属与结构。方法 建立大鼠ig给予XYP-A后血清中移行成分的UPLC-PDA色谱分析方法。对全方石油醚萃取物及柴胡（红柴胡）、当归、白术各单方石油醚萃取物分别ig给予大鼠后血清样品色谱图进行比较,分析血清中移行成分的归属,通过与对照品比对,比较保留时间、UV扫描图并结合文献数据对其定性。结果 大鼠ig给予XYP-A后,血清中共检测到20个入血成分,其中12个为原形成分,分别来源于柴胡9个、当归2个、白术1个,另外8个为代谢产物,并指认了4个原形成分,分别为藁本内酯、白术内酯II、2, 8, 10-十五烷三烯-4, 6-二炔-1-醇和柴胡炔醇。结论 所建立的方法可行,血中检测到的20个移行成分很可能是逍遥散体内直接作用物质,为阐明该方药效物质基础提供了依据。